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WSP-1 (Synonyms: Washington State Probe1)

Catalog No.GC45161

WSP-1 is a selective and rapid responsive fluorescent probe for active sulfides, specifically designed for the detection of hydrogen sulfide (H2S) within biological samples and intracellular environments.

Products are for research use only. Not for human use. We do not sell to patients.

WSP-1 Chemical Structure

Cas No.: 1352750-34-5

Size Price Stock Qty
1mg
$56.00
In stock
5mg
$185.00
In stock
10mg
$339.00
In stock

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Sample solution is provided at 25 µL, 10mM.

Product Documents

Quality Control & SDS

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Protocol

WSP-1 Usage Instructions:

Preparation of Stock Solution: Remove the product from the refrigerator and allow it to equilibrate at room temperature. Centrifuge at low speed for 3-5 minutes. Add an appropriate amount of anhydrous DMSO (Dimethyl Sulfoxide) to the vial and dissolve thoroughly to prepare a 5mM stock solution. Divide the solution into single-use aliquots and store them in the freezer, avoiding repeated freeze-thaw cycles. The stock solution should remain stable for at least three months.

For specific working concentrations, please refer to the literature or adjust as needed based on your experimental requirements.

Protocol for for Fluorescent Detection of Intracellular H2S (Fluorescence Microscopy Examination) by WSP-1 [1]:

  1. 1.Culture HASMCs until they reach approximately 90% confluence. 24 hours before the experiment, seed the cells on 8-well culture slides at a density of 30,000 cells per well and incubate for 24 hours.
  2. 2.After 24 hours, allow the cells to adhere, replace the culture medium with 400 μL of WSP-1(100 μM)working solution, and incubate at 37℃ for 30 minutes.
  3. 3.Replace the WSP-1 working solution with 380 μL of standard buffer and add 20 μL of the required concentration of H2S donor compound.
  4. 4.When WSP-1 reacts with hydrogen sulfide, a detectable fluorescence signal is released at Ex/Em = 465/515 nm. After incubating for 1 hour, wash the cells with 400 μL of DPBS.
  5. 5.Remove the DPBS, and at room temperature, add 200 μL of Bouin's solution to each well to fix the cells for 10 minutes. Remove excess Bouin's solution and wash the cells twice with 400 μL of DPBS.
  6. 6.Use a fluorescence microscope to examine and capture the fluorescence signals.

This protocol only provides a guideline, and should be modified according to your specific needs.

Protocol for Visualizing Intracellular H2S Using WSP-1 [2]:

  1. 1.The roots of seedlings after treatments were transferred to 20 mM Hepes-NaOH (pH 7.5) buffer solution containing 15 μM of WSP-1.
  2. 2.After being incubated in darkness at 25℃ for 40 min, the roots were washed with distilled water three times and were visualized immediately by a fluorescence microscope with a 465/515 nm and an excitation/emission filter set.
  3. 3.For the detection of WSP-1 fluorescence in different reactive sulfur species, WSP-1 with final concentration of 15 μM were added into the following solutions, SDS (sodium dodecyl sulfate, 2 mM), NaHSO4 (2 mM), NaHSO3 (2 mM), Na2SO4 (2 mM), Na2SO3 (2 mM), Na2S2O4 (2 mM), GSSG (glutathione disulfide, 2 mM), sulfonamide (2 mM), GSNO (2 mM), and NaHS (2 mM), respectively. 20 μL of the above solutions were transferred to a glass slide for the visualization with a fluorescence microscope with a 465/515 nm and an excitation/emission filter set.

This protocol only provides a guideline, and should be modified according to your specific needs.

References:

[1]. Martelli A, Citi V,et,al. Vascular Effects of H2S-Donors: Fluorimetric Detection of H2S Generation and Ion Channel Activation in Human Aortic Smooth Muscle Cells. Methods Mol Biol. 2019;2007:79-87. doi: 10.1007/978-1-4939-9528-8_6. PMID: 31148107.

[2]. Li YJ, Chen J, et,al. In site bioimaging of hydrogen sulfide uncovers its pivotal role in regulating nitric oxide-induced lateral root formation. PLoS One. 2014 Feb 27;9(2):e90340. PMID: 24587333

Background

WSP-1 is a selective and rapid responsive fluorescent probe for active sulfides, specifically designed for the detection of hydrogen sulfide (H2S) within biological samples and intracellular environments. WSP-1 selectively and rapidly reacts with H2S, generating benzodithiolone and a fluorescent compound (Ex=465nm, Em=515nm) [1].

WSP-1 (10 µM) can detect sulfide levels in RFL-6 cells where the sulfide donor GYY4137 (10-100 µM) is present[2]. The use of the WSP-1 fluorescent dye enables the detection of hydrogen sulfide (H2S) generation in human vascular smooth muscle cells following H2S donor administration[3]. WSP-1 can applied to track endogenous H2S in tomato (Solanum lycopersicum) roots in site[4].

References:
[1]. Liu C, Pan J, et,al. Capture and visualization of hydrogen sulfide by a fluorescent probe. Angew Chem Int Ed Engl. 2011 Oct 24;50(44):10327-9. doi: 10.1002/anie.201104305. Epub 2011 Sep 6. PMID: 21898737; PMCID: PMC3417056.
[2]. Cortese-Krott MM, Fernandez BO, et,al. Nitrosopersulfide (SSNO(-)) accounts for sustained NO bioactivity of S-nitrosothiols following reaction with sulfide. Redox Biol. 2014 Jan 11;2:234-44. doi: 10.1016/j.redox.2013.12.031. PMID: 24494198; PMCID: PMC3909780.
[3]. Martelli A, Citi V,et,al. Vascular Effects of H2S-Donors: Fluorimetric Detection of H2S Generation and Ion Channel Activation in Human Aortic Smooth Muscle Cells. Methods Mol Biol. 2019;2007:79-87. doi: 10.1007/978-1-4939-9528-8_6. PMID: 31148107.
[4]. Li YJ, Chen J, et,al. In site bioimaging of hydrogen sulfide uncovers its pivotal role in regulating nitric oxide-induced lateral root formation. PLoS One. 2014 Feb 27;9(2):e90340. doi: 10.1371/journal.pone.0090340. PMID: 24587333; PMCID: PMC3937356.

Chemical Properties

Cas No. 1352750-34-5 SDF
Synonyms Washington State Probe1
Chemical Name 3'-methoxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthen]-6'-yl 2-(pyridin-2-yldisulfanyl)benzoate
Canonical SMILES O=C(OC1=CC=C2C(OC3=C(C24C5=CC=CC=C5C(O4)=O)C=CC=C3)=C1)CO=C(OC1=CC=C2C(OC3=C(C24C5=CC=CC=C5C(O4)=O)C=CC(OC)=C3)=C1)C6=C(SSC7=NC=CC=C7)C=CC=C6
Formula C33H21NO6S2 M.Wt 591.7
Solubility 2mg/mL in DMSO, 20mg/mL in DMF Storage Store at -20°C, protect from light
General tips Please select the appropriate solvent to prepare the stock solution according to the solubility of the product in different solvents; once the solution is prepared, please store it in separate packages to avoid product failure caused by repeated freezing and thawing.Storage method and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored at -20°C, please use it within 1 month.
To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time.
Shipping Condition Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table

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1 mg 5 mg 10 mg
1 mM 1.69 mL 8.4502 mL 16.9005 mL
5 mM 0.338 mL 1.69 mL 3.3801 mL
10 mM 0.169 mL 0.845 mL 1.69 mL
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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.

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